苯基吡啶酮衍生物影响猪δ冠状病毒复制的作用分析

doi:10.11843/j.issn.0366-6964.2022.09.033

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (9): 3190-3198.doi: 10.11843/j.issn.0366-6964.2022.09.033

苯基吡啶酮衍生物影响猪δ冠状病毒复制的作用分析

姚晨^1,2, 郭萌^1,3, 胡慧^1,3, 史晨曦^1,3*, 杨国宇^1,2

1. 河南农业大学动物医学院, 郑州 450002;
2. 河南农业大学农业农村部动物生化与营养重点实验室, 郑州 450002;
3. 河南省动物性食品安全重点实验室, 郑州 450002

收稿日期:2022-01-28 出版日期:2022-09-23 发布日期:2022-09-23
通讯作者: 史晨曦,主要从事动物生物化学研究,E-mail:shichenxi@henau.edu.cn
作者简介:姚晨(1998-),男,河南周口人,硕士生,主要从事动物生物化学研究,E-mail:873932556@qq.com
基金资助:
河南省杰出青年基金项目（202300410192）

Effects of Phenylpyridinone Derivative on the Replication of Porcine Deltacoronavirus

YAO Chen^1,2, GUO Meng^1,3, HU Hui^1,3, SHI Chenxi^1,3*, YANG Guoyu^1,2

1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;
2. Key Laboratory of Animal Biochemistry and Nutrition of Ministry of Agriculture and Rural Affairs, Henan Agricultural University, Zhengzhou 450002, China;
3. Henan Province Key Laboratory of Animal Food Safety, Zhengzhou 450002, China

Received:2022-01-28 Online:2022-09-23 Published:2022-09-23

摘要/Abstract

摘要： 本研究旨在探讨苯基吡啶酮衍生物JIB-04对猪δ冠状病毒（porcine deltacoronavirus，PDCoV）的抗病毒作用，并初步研究可能的作用机制。首先应用CCK-8方法检测不同浓度JIB-04作用后的细胞活力，并计算半数毒性浓度（CC₅₀）和半数有效浓度（EC₅₀）。应用TCID₅₀方法检测JIB-04预处理和共处理对PDCoV复制的影响，检测JIB-04处理对PDCoV吸附或入侵细胞的影响。最后，应用qRT-PCR、TCID₅₀和Western blot方法检测JIB-04处理对不同时间点病毒复制的影响。结果表明，JIB-04在所有受试浓度下均不影响LLC-PK细胞的活力，CC₅₀>640 μmol·L^-1，EC₅₀=0.216 μmol·L^-1，SI指数>2 963。与未处理的病毒感染组相比，JIB-04处理极显著降低了PDCoV的滴度（P<0.001），但对PDCoV吸附和入侵过程没有影响。感染6 h后，与未处理的病毒感染组相比，JIB-04处理组PDCoV滴度极显著下降（P<0.01）；感染12和24 h后，PDCoV滴度、基因组拷贝数、N蛋白表达水平均极显著下降（P<0.01）。JIB-04的细胞毒性较低，药物选择性指数较高，在体外能抵抗PDCoV感染，可以作为潜在的抗病毒药物。在PDCoV感染过程中，JIB-04能够抑制病毒核酸和蛋白质的合成，抑制病毒的复制。

关键词: 苯基吡啶酮衍生物JIB-04, 抗病毒, 猪δ冠状病毒

Abstract: The purpose of this study was to investigate the antiviral effect of phenylpyridinone derivative JIB-04 on porcine deltacoronavirus (PDCoV), and explore the possible mechanism. Cell viability after treatment with different concentrations of JIB-04 was detected by CCK-8, and the 50% cytotoxic concentration (CC₅₀) and 50% effective concentration (EC₅₀) were calculated. TCID₅₀ method was used to detect the effects of JIB-04 pretreatment and co-treatment on PDCoV replicationand the effect of JIB-04 treatment on virus attachment and penetration. Finally, qRT-PCR, TCID₅₀ and Western blot methods were used to detect the effect of JIB-04 on virus replication at different times post infection. The results showed that JIB-04 did not affect the cell viability of LLC-PK cells at all tested concentrations, and CC₅₀>640 μmol·L^-1, EC₅₀=0.216 μmol·L^-1, and SI index is greater than 2 963. Compared with untreated virus infection group, JIB-04 treatment significantly reduced the virus titer (P<0.001), but it had no effect on attachment or penetration of PDCoV. At 6 h post infection, compared with untreated virus infection group, virus titer in JIB-04 treatment group was significantly decreased (P<0.01). At 12 and 24 h post infection, virus titer, genome copy number, and N protein expression level all significantly decreased (P<0.01). JIB-04 has a low cytotoxicity and a high selective index, and can protect against PDCoV infection in vitro, making it a potential antiviral drug. JIB-04 can inhibit synthesis of viral RNA, protein and PDCoV replication.

Key words: phenylpyridinone derivative JIB-04, antiviral, porcine deltacoronavirus

中图分类号:

R978.7

姚晨, 郭萌, 胡慧, 史晨曦, 杨国宇. 苯基吡啶酮衍生物影响猪δ冠状病毒复制的作用分析[J]. 畜牧兽医学报, 2022, 53(9): 3190-3198.

YAO Chen, GUO Meng, HU Hui, SHI Chenxi, YANG Guoyu. Effects of Phenylpyridinone Derivative on the Replication of Porcine Deltacoronavirus[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3190-3198.

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苯基吡啶酮衍生物影响猪δ冠状病毒复制的作用分析

Effects of Phenylpyridinone Derivative on the Replication of Porcine Deltacoronavirus

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